A Novel Triazolopyridine-Based Spleen Tyrosine Kinase Inhibitor That Arrests Joint Inflammation.

Autoantibodies and the immunoreceptors to which they bind can contribute to the pathogenesis of autoimmune diseases such as rheumatoid arthritis (RA). Spleen Tyrosine Kinase (Syk) is a non-receptor tyrosine kinase with a central role in immunoreceptor (FcR) signaling and immune cell functionality. Syk kinase inhibitors have activity in antibody-dependent immune cell activation assays, in preclinical models of arthritis, and have progressed into clinical trials for RA and other autoimmune diseases. Here we describe the characterization of a novel triazolopyridine-based Syk kinase inhibitor, CC-509. This compound is a potent inhibitor of purified Syk enzyme, FcR-dependent and FcR-independent signaling in primary immune cells, and basophil activation in human whole blood. CC-509 is moderately selective across the kinome and against other non-kinase enzymes or receptors. Importantly, CC-509 was optimized away from and has modest activity against cellular KDR and Jak2, kinases that when inhibited in a preclinical and clinical setting may promote hypertension and neutropenia, respectively. In addition, CC-509 is orally bioavailable and displays dose-dependent efficacy in two rodent models of immune-inflammatory disease. In passive cutaneous anaphylaxis (PCA), CC-509 significantly inhibited skin edema. Moreover, CC-509 significantly reduced paw swelling and the tissue levels of pro-inflammatory cytokines RANTES and MIP-1α in the collagen-induced arthritis (CIA) model. In summary, CC-509 is a potent, moderately selective, and efficacious inhibitor of Syk that has a differentiated profile when compared to other Syk compounds that have progressed into the clinic for RA.

IKK-i signals through IRF3 and NFkappaB to mediate the production of inflammatory cytokines.

IKK-i and TBK1 were recently identified as IKK-related kinases that are activated by toll-like receptors TLR3 and TLR4. These kinases were identified as essential components of the virus-activated as well as LPS-MyD88 independent kinase complex that phosphorylates IRF3 and results in the production of cytokines involved in innate immunity. Both IKK-i and TBK1 have also been implicated in the activation of the NFkappaB pathway but the precise mechanism is not clear. Although the literature to date suggests that IKK-i and TBK1 play redundant roles in TLR3 and TLR4 signaling, recent data suggest that there may be subtle differences in the signaling pathways affected by these kinases. We have generated tetracycline-inducible stable cell lines that express a wild type or kinase-inactive mutant form of IKK-i. Our data suggest that expression of IKK-i can activate both NFkappaB and IRF3, leading to the production of several cytokines including interferon beta. IKK-i most likely acts upstream of IKK2 to activate NFkappaB in these cells since expression of the kinase-inactive version of IKK-i did not inhibit TNFalpha mediated production of inflammatory cytokines. The data suggest that IKK-i is not involved in TNF-alpha mediated signaling but instead could likely play a role in activating IKK2 downstream of Toll-like receptor signaling. We also identified STAT1, Tyk2, and JAK1 as secondary mediators of IKK-i signaling as a result of interferon beta production in these cells.

Comparison of four diagnostic methods for detection of Helicobacter species in laboratory mice.

Four species of Helicobacter--H. muridarum, "H. rappini," H. hepaticus, and H. bilis--have been identified in the gastrointestinal tract of rodents. The association of Helicobacter species with chronic gastrointestinal diseases in mice has raised concern about their impact on research results. In this study, different methods for detection of Helicobacter species in the mouse intestinal tract were compared: polymerase chain reaction (PCR) amplification of 16S rRNA gene sequences, bacterial culture, electron microscopy, and histologic examination (Steiner stain). The PCR method was more sensitive in detecting murine Helicobacter species than was culture, electron microscopy, or histologic examination. Of the cecal specimens identified as Helicobacter species-positive by PCR, approximately 60% were identified as positive by each of the other methods. An 87.5% concordance was obtained by PCR screening of DNA from fecal and cecal specimens. Differentiation among murine Helicobacter species by colony morphologic or histologic features was not possible. Scanning electron microscopy and histologic examination indicated greater numbers of helical microorganisms, specifically H. hepaticus, in the cecum than in the colon. These results indicate that the PCR assays used can be performed on feces as a noninvasive means for rapidly screening large numbers of colony mice for murine Helicobacter species.

Immunohistochemical identification of prostatic acid phosphatase: correlation of tumor grade with acid phosphatase distribution.

The localization and distribution of prostatic specific acid phosphatase (PSAP) in normal, hyperplastic and neoplastic prostates were studied by specific immunohistochemical of normal and hyperplastic prostates. In adenocarcinoma of the prostate, a correlation of the PSAP staining with the degree of differentiation and the ability of the tumor to form a gland was observed: more intense and uniform staining in well differentiated tumors and less intense and more variable stains in poorly differentiated tumors. The same correlation was also observed in tumors metastasized to lymph nodes and other organs.

Segmental calculus disease: potential of parital nephrectomy.

Partial nephrectomy was performed on 71 renal units between 1962 and 1978 for segmental calculus disease. Parenchymal scarring associated with an infundibulocalyceal stone, which was usually branched, was the indication for resection. Stone analysis demonstrated an equal incidence of idiopathic and struvite stones. Perioperative morbidity was minimal, but pyelocutaneous urinary drainage prolonged the hospitalization of 5 patients. In 2 cases, the cause was an obstructing retained calculus. Retained calculi occurred in 3 other patients, one requiring early nephrectomy for sepsis. Fifty-seven patients were followed for longer than twelve months. Ipsilateral calculi recurred in 12 per cent of kidneys, and contralateral new calculi developed in the same number. Ninety-four per cent of patients with preoperative urinary tract infections had sterile urine at follow-up. From the results of this and other series, partial nephrectomy compares favorably with extended pyelolithotomy and anatrophic nephrotomy for segmental calculus disease associated with parenchymal scarring and/or a deformed collecting system.

Guidelines in the management of patients with pancreatic abscess.

A systematic approach to the management of patients with pancreatic abscess is presented. The method includes an aggressive diagnostic approach relying on radiographic and ultrasound examinations, adequate transperitoneal drainage of the abscess, coupled with gastrostomy and feeding jejunostomy. The importance of nutritional support using an elemental diet administered through a jejunostomy is emphasized.

Cardiac rupture--challenge in diagnosis and management.

Previous studies of the incidence, natural history, pathogenesis and diagnosis of cardiac rupture are presented, and 20 additional cases described. Progressive cardiorrhexis after myocardial infarction causes death in possibly more than 25,000 persons a year in the United States, and more frequent antemortem diagnosis is needed. Suggestions for future clinical and experimental studies are described and possible means of early diagnosis and therapy are outlined.

Acute aortic arch dissection: reevaluation of the indications for medical and surgical therapy.

Of 42 patients with dissection of the aorta, 4 had important arch involvement. Results were good in 2 patients treated medically. In two other patients wrapping the arch with a Dacron graft successfully prevented fatal hemorrhage. This technique avoids the need for arch replacement in selected cases. From this experience and a review of others a flow sheet was developed to guide decision-making in the surgical and medical management of patients with aortic dissection.

The cardiac veins and retrograde coronary venous perfusion.

Study of retrograde coronary venous perfusion has been greatly overshadowed in the past decade by the advent and success of direct coronary artery revascularization. Recently there has been renewed interest in retrograde venous perfusion as a possible means of restoring myocardial circulation in selected patients. This paper reviews the anatomy and physiology of the cardiac venous system and the history of retrograde venous perfusion, with emphasis on recent experimental work and clinical trials involving retrograde coronary vein perfusion (RCVP).